Attachment and Proliferation of Retinal Pigment Epithelial Cells on Small Intestine Submucosa Powder Impregnated Poly(l-Lactide-Co-Glycolide) Film

Abstract

The retinal pigment epithelium (RPE) closely interacts with photoreceptors in the maintenance of visual function. The native RPEs exist as a monolayer structure and have a mottled brown color due to the presence of melanin and other pigments including lipofuscin granules, which accumulate with age. In age-related macular degeneration (AMD), RPE's dysfunction and changes in Bruch's membrane occur. Thus, small intestine submucosa/poly(lactic-co-glycolic acid) (SIS/PLGA) film is a biomimetic transplant consisting of a layer of healthy RPE cells cultured on a support membrane. The goals of this study were to evaluate the effects of attachment and proliferation of RPEs on SIS/PLGA films. Porcine SIS is an acellular tissue and widely used as a biomaterial without immunorejection responses, whereas PLGA is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. We fabricated SIS/PLGA films using 20 wt% of SIS. We measured MTT to confirm cellular adhesion of cell number attached on film at 1, 3, 5, and 7 days. Morphology of cellular adhesion on films was confirmed by scanning electron microscopy at 1, 3, and 7 days. Further, reverse transcription polymerase chain reaction (RT-PCR) was conducted to confirm messenger RNA expression of RPE65 as RPE's marker and expression of cytokeratin, and RPE65 were determined by AEC immunocytochemical staining. These results suggest that SIS provides suitable surface to RPEs.

Effects of small intestinal submucosa content on the adhesion and proliferation of retinal pigment epithelial cells on SIS-PLGA films

“…The reaction products were then mixed with avidin–biotin peroxidase (Immunotech) and reacted for another 30 min at room temperature. The colour change was evaluated with a chromogen (Immunotech) and examined by light microscopy (Jo et al ., ).…”